Andrea Vanesa Toledo

Histopathology Caused by the Entomopathogenic Fungi, Beauveria bassiana and Metarhizium anisopliae, in the Adult Planthopper, Peregrinus maidis, a Maize Virus Vector

Abstract The planthopper Peregrinus maidis (Ashmead) (Hemiptera: Delphacidae) is an important vector of maize viruses in tropical and subtropical areas. Planthoppers are biologically controlled with several species of entomopathogenic fungi that have been isolated from these insect pests of rice in Asia. Beauveria bassiana (Balsamo-Crivelli) Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin (Hypocreales: Clavicipitaceae) appear to be the most useful against planthoppers because of their ease of mass production, storage, virulence, and application. In the present study, adults of P. maidis infected with B. bassiana and M. anisopliae were observed under light and scanning electron microscopy to characterize morphologically the process of infection and the development of these fungi, prior to and after the death of the host. The hydrophobic conidia of both fungal species were able to attach to all body regions, with a preference for surfaces containing hairs. Few germinated conidia were observed on the insects body surface at 24, 48, and 72 hr post-inoculation. On the cuticular surface of P. maidis treated with B. bassiana and M. anisopliae, bacillus-like bacteria were observed. These microorganisms could be interacting with fungal conidia, playing a role of antibiosis that will not allow the fungal pathogens to germinate and penetrate. In the colonization events observed in this study, the formation and multiplication of hyphal bodies by both fungal species inside the hosts body was noted. The hosts whole body was invaded by hyphae between five and six days post-inoculation, and body fat was the most affected tissue.

Growth Inhibition of Beauveria bassiana by Bacteria Isolated from the Cuticular Surface of the Corn Leafhopper, Dalbulus maidis and the Planthopper, Delphacodes kuscheli, Two Important Vectors of Maize Pathogens

Abstract The phytosanitary importance of the corn leafhopper, Dalbulus maidis (De Long and Wolcott) (Hemiptera: Cicadellidae) and the planthopper, Delphacodes kuscheli Fennah (Hemiptera: Delphacidae) lies in their ability to transmit phloem-associated plant pathogens, mainly viruses and mollicutes, and to cause considerable mechanical damage to corn plants during feeding and oviposition. Fungi, particularly some members of the Ascomycota, are likely candidates for biocontrol agents against these insect pests, but several studies revealed their failure to invade the insect cuticle possibly because of the presence of inhibitory compounds such as phenols, quinones, and lipids and also by the antibiosis effect of the microbiota living on the cuticular surface of the host. The present work aims to understand interactions between the entomopathogenic fungus Beauveria bassiana (Balsamao-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae) and bacterial antagonists isolated from the cuticular surface of D. maidis and D. kuscheli. A total of 155 bacterial isolates were recovered from the insects cuticle and tested against B. bassiana. Ninety-one out of 155 strains inhibited the growth of B. bassiana. Bacterial strains isolated from D. maidis were significantly more antagonistic against B. bassiana than those isolates from D. kuscheli. Among the most effective antagonistic strains, six isolates of Bacillus thuringiensis Berliner (Bacillales: Bacillaeae (after B. subtilis)), one isolate of B. mycoides Flügge, eight isolates of B. megaterium de Bary, five isolates of B.pumilus Meyer and Gottheil, one isolate of B. licheniformis (Weigmann) Chester, and four isolates of B. subtilis (Ehrenberg) Cohn were identified.

Pathogenic fungi of planthoppers associated with rice crops in Argentina

We conducted a survey of pathogenic fungi of planthoppers associated with rice crops in Los Hornos, Buenos Aires, Argentina, and a study of the seasonality and prevalence of these pathogens. Samples were taken in rice for two consecutive years. The plants sampled included rice Oryza sativa L. (Poaceae) and its surrounding weeds. The planthopper Oliarus dimidiatus Berg. (Hemiptera: Fulgoromorpha: Cixiidae) was the most abundant species associated with rice. Two species of entomophthoralean and one species of hypocrealean fungi infected and killed adults of O. dimidiatus in the rice agroecosystem. The fungi were identified as: Pandora sp. (Entomophthorales: Entomophthoraceae), Conidiobolus coronatus (Costantin) Batko (Entomophthorales: Ancylistaceae), and Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Clavicipitaceae). Pandora sp. was the most predominant pathogen collected. It was recorded from the middle of February to the middle of April 2005, and not found again until the end of March 2006. The report of Pandora sp., C. coronatus and B. bassiana represents the first records of these fungi as pathogens of hoppers associated with rice crops in Argentina.

Morphological and molecular characterization of a fungus, Hirsutella sp., isolated from planthoppers and psocids in Argentina

Abstract A mycosed planthopper, Oliarus dimidiatus Berg (Hemiptera: Cixiidae), and two psocids, Heterocaecilius sp. (Psocodea: Pseudocaeciliidae) and Ectopsocus sp. (Ectopsocidae), were collected from Los Hornos and La Plata, Buenos Aires, Argentina between February and September 2007. Observations of mycelia growing on the host revealed that the putative fungal parasite had synnemata supporting monophialidic conidiogenous cells. Likewise, in vitro fungal cultures presented characteristics typical of the fungus Hirsutella citriformis Speare (Ascomycota: Hypocreales: Clavicipitaceae). The identity of the isolated fungi characterized based on morphological aspects was complemented by means of the internal transcribed spacer sequences. The sequences of both isolates were highly homologous to those of Cordyceps sp. (Fries) Link and Ophiocordyceps sinensis (Berkely) G.H. Sung, J.M. Sung, Hywel-Jones, and Spatafora (Ophiocordycipitaceae). We additionally confirmed that both isolates had the ability to infect and kill adults of Delphacodes kuscheli Fennah (Hemiptera: Delphacidae) after 10 days. Therefore, based on the morphology of the isolated fungi, their ribosomal internal transcribed spacer sequence, and their ability to parasite insects, we conclude that the fungi isolated belong to the genus Hirsutella and might have biotechnological potential.

Antagonism of entomopathogenic fungi by Bacillus spp. associated with the integument of cicadellids and delphacids

Entomopathogenic fungi are potential tools to biocontrol cicadellids and delphacids, two groups of insects that cause extensive damage to agricultural crops. However, bacteria living on the host cuticle may inhibit fungal growth. In the present work, following the molecular characterization of 10 strains of Bacillus isolated from the integument of cicadellids and delphacids, we selected isolates of the fungi Beauveria bassiana and Metarhizium anisopliae that are resistant to the antimicrobials secreted by these bacterial strains. The antagonistic activity of the 10 bacterial isolates belonging to the genus Bacillus (i.e., B. amyloliquefaciens, B. pumilus, and B. subtilis) against 41 isolates of Bea. bassiana and 20 isolates of M. anisopliae was investigated in vitro on tryptic soy agar using the central disk test. With this approach, isolates of Bea. bassiana and M. anisopliae resistant to antagonistic bacteria were identified that can be further developed as biological control agents.

Description of the immature stages of Delphacodes kuscheli Fennah (Hemiptera: Delphacidae), vector of “Mal de Río Cuarto virus” on maize in Argentina

Delphacodes kuscheli Fennah is the most efficient vector of Mal de Río Cuarto virus (MRCV) maize disease in Argentina, which is caused by a persistently transmitted fijivirus. Delphacodes kuscheli is abundant in a wide area of Central Argentina (32–35°S), affecting oat, wheat and maize crops, and weeds. All five immature instars of D. kuscheli are described, keyed and illustrated. A key is provided to highlight the main features for identification of the fifth‐instar nymphs of three other Delphacid species on maize. Adults of D. kuscheli were reared under controlled conditions in individual glass cages containing oat plants. The description of each instar was based on 1‐day‐old nymphs. Coloration pattern, shape of frons, length of the rostrum segments, and length, shape and number of spur teeth are useful features for identification of the fifth nymphal instars of these species.

Diversity of planthoppers (Hemiptera: Fulgoromorpha) in rice associated with weeds in Argentina

An assemblage of Fulgoromorpha in a rice agroecosystem from La Plata city was studied for the first time in Argentina, considering those species associated with rice and the surrounding weeds, their seasonality, sex ratios and parasitoidism. Samples were obtained from 2006 to 2010 in a rice field and in the surrounding weeds. Fourteen species from three families of Fulgoromorpha were associated with rice, most of them native species. The delphacids Chionomus haywardi, Toya propinqua and Delphacodes kuscheli were the most abundant on rice. Population peaks occurred after flooding of rice and before anthesis. The native C. haywardi was more abundant in rice than in weeds, while the opposite was confirmed for T. propinqua, a well-known cosmopolitan grass specialist. The difference of abundances of D. kuscheli in rice and weeds was not significant. Our results highlight the dominance of C. haywardi during all sampling years in the rice agroecosystem as well as the role of surrounding weeds as an alternative microhabitat for planthoppers instead of rice. The strepsipteran Elenchus tenuicornis was the only recorded parasitoid and showed no preference for any of the insect hosts.